General Cleaning and Maintenance of the Laboratory

The introduction of a cleaning routine into the tissue culture laboratory should result in
lessening the risk of contamination, maintenance of researcher’s safety, and fewer
problems with cell growth.

1. Check incubator temperatures and CO2 levels daily

2. Wipe down bench surfaces with 70% alcohol
3. Wipe the working surface of the laminar flow hood with 70% alcohol
4. If using a vacuum, check the pump is functioning and that the aspirator is clean
and contains Clorox
5. Any media bottles, etc. should be wiped with 70% alcohol before placing in hood
6. Water baths should be cleaned regularly and replaced with fresh water
7. The use of Bunsen burners in hoods is not recommended as the flame disrupts the
air flow
8. After use of hood, turn off vacuum pump, discard any waste into suitable bags for
autoclaving, check there is sufficient Clorox in waste flask and the flask is not
over-full, and wipe interior with 70% alcohol. Turn on UV for a short time
9. Cell cultures should be observed microscopically daily for one to become familiar
with the morphological appearance of different cells. This will lead to an ability to
see when irregularities and contamination occur which, in turn, may allow one to
rescue or discard a culture at an early stage
10. At the end of the day remember to switch off microscopes, turn off water baths,
etc. and check to make sure incubator doors are closed, and lids on LN2 tanks are
replaced properly
11. Clean all hoods thoroughly, removing the working surface tray so that the base is
accessible. Spillages can easily get through to the base. Clean with a bacdown
solution and wipe with 70% alcohol
12. Clean out all incubators regularly and thoroughly, removing all shelving and
paying particular attention to door seals. If a tray is been used to hold water for
humidification, this should be removed, cleaned and refilled with fresh sterile
water. Roccal (Benzalkonium chloride- from Sigma Aldrich) can be added to the
tray at the specified concentration (1:64 from a 10% solution)
13. Check incubator water levels before the weekend and top-up if necessary
14. Check HEPA filter inside the incubator- replace every 6months.
15. Cell stocks should not be in culture for more than three months. Recover a new
vial and make sure cells are tested regularly for Mycoplasma. If cells are
contaminated with mycoplasma it is wise to discard cultures. If that is not possible
cells can be treated with antibiotics such as MRA (Mycoplasma Removal Agent)
or Plasmocin. Double the treatment time to ensure mycoplasma is completely
gone

Carmel Burns Page 1 7/13/2019